Stimulation of multipotency of mesenchymal stem cells by chemokine ccl5

Abstract

Methods of stimulating multipotency, proliferation and differentiation of isolated mesenchymal stem cells (MSCs), which permit more effective differentiation and integration of such cells into host tissues. The method includes providing an in vitro cell population of MSCs and administering CCL5 chemokine. Preferably CCL is administered in an amount sufficient to induce expression of one or more multipotency related genes selected from the group consisting of OCT3/4, NANOG, SOX 2, KLF4, and SOX9.

What is claimed is: 
     
         1 . A method of increasing expression of a chemokine receptor in a population of mesenchymal stem cells comprising:
 a) providing an in vitro population of human mesenchymal stem cells; and   b) administering CCL5 to the population in an amount sufficient to increase expression of a chemokine receptor selected from the group consisting of CCR1, CCR2 and CCR5 in the population.   
     
     
         2 . The method according to  claim 1 , wherein the method increases expression of CCR1, CCR3 and CCR5 in the population of cells. 
     
     
         3 . The method according to  claim 1 , further comprising culturing the population of cells for about 48 hours after the step of administering CCL5 to achieve a higher level of expression of the chemokine receptor compared to a same population of cells cultured for about 24 hours and for about 72 hours after the step of administering CCL5. 
     
     
         4 . A method of increasing expression of a NFkB-dependent gene selected from the group consisting of IL-6, MMP1, and MMP9 in a population of mesenchymal stem cells, the method comprising:
 a) providing a in vitro population of human mesenchymal stem-like cells; and   b) administering CCL5 to the population in an amount sufficient to increase expression of a NFkB-dependent gene selected from the group consisting of IL-6, MMP1, and MMP9.   
     
     
         5 . The method according to  claim 4 , wherein expression of an anti-apoptotic gene SURVIVINE is increased. 
     
     
         6 . The method according to  claim 5 , the method further comprising culturing the population of cells for about 8 hours after administering CCL5 to obtain a higher level of expression of IL-6, MMP1, MMP9 and SURVIVINE and culturing the population of cells for about 24 hours to obtain a higher level of expression of MMP2. 
     
     
         7 . A method of inducing expression of multipotency related genes selected from the group consisting of OCT3/4, NANOG, SOX 2, KLF4, AND SOX9 in mesenchymal stem cells, the method comprising:
 a) providing a in vitro population of human mesenchymal stem cells; and   b) administering CCL5 to the population in an amount sufficient to induce expression of one or more multipotency related genes selected from the group consisting of OCT3/4, NANOG, SOX 2, KLF4, and SOX9.   
     
     
         8 . The method according to  claim 7 , further comprising the step of culturing the population for about 48 hours from the step of administering CCL5 to achieve a higher level of expression of OCT 3/4, NANOG, and SOX2 compared to about 8 hours, 24 hours and 72 hours after the step of administering CCL5. 
     
     
         9 . A method of stimulating proliferation of mesenchymal stem cells comprising:
 a) providing an in vitro population of human mesenchymal stem cells; and   b) administering CCL5 to the population in an amount sufficient to stimulate proliferation.   
     
     
         10 . A method stimulating differentiation of mesenchymal stem cells, comprising:
 a) providing an in vitro population of human mesenchymal stem cells; and   b) administering CCL5 to the population in an amount sufficient to stimulate differentiation of mesenchymal stem cells.   
     
     
         11 . The method according to  claim 10 , wherein the population of cells is cultured for 12 hours with CCL5 followed by differentiation into adipogenic or chondrogenic lineages. 
     
     
         12 . The method according to  claim 11 , further comprising determining expression levels of PPARγ2, ADIPOQ, CFD and FABP4 to confirm an adipogenic and lineage. 
     
     
         13 . The method according to  claim 11 , further comprising determining expression levels of SOX9, COL2A1, NKX3.1 and MMP13 to confirm a chondrogenic lineage.